全文获取类型
收费全文 | 5823篇 |
免费 | 388篇 |
国内免费 | 34篇 |
出版年
2023年 | 15篇 |
2022年 | 45篇 |
2021年 | 76篇 |
2020年 | 44篇 |
2019年 | 60篇 |
2018年 | 64篇 |
2017年 | 71篇 |
2016年 | 110篇 |
2015年 | 178篇 |
2014年 | 204篇 |
2013年 | 400篇 |
2012年 | 344篇 |
2011年 | 359篇 |
2010年 | 200篇 |
2009年 | 197篇 |
2008年 | 357篇 |
2007年 | 357篇 |
2006年 | 367篇 |
2005年 | 360篇 |
2004年 | 338篇 |
2003年 | 346篇 |
2002年 | 309篇 |
2001年 | 140篇 |
2000年 | 148篇 |
1999年 | 99篇 |
1998年 | 96篇 |
1997年 | 78篇 |
1996年 | 47篇 |
1995年 | 38篇 |
1994年 | 33篇 |
1993年 | 51篇 |
1992年 | 73篇 |
1991年 | 52篇 |
1990年 | 56篇 |
1989年 | 46篇 |
1988年 | 46篇 |
1987年 | 28篇 |
1986年 | 38篇 |
1985年 | 33篇 |
1984年 | 30篇 |
1983年 | 36篇 |
1982年 | 31篇 |
1981年 | 23篇 |
1980年 | 23篇 |
1979年 | 25篇 |
1978年 | 25篇 |
1977年 | 21篇 |
1974年 | 14篇 |
1970年 | 17篇 |
1968年 | 15篇 |
排序方式: 共有6245条查询结果,搜索用时 15 毫秒
61.
Details of Retropositional Genome Dynamics That Provide a Rationale for a Generic Division: The Distinct Branching of All the Pacific Salmon and Trout (Oncorhynchus) from the Atlantic Salmon and Trout (Salmo) 总被引:2,自引:2,他引:0
下载免费PDF全文
![点击此处可从《Genetics》网站下载免费的PDF全文](/ch/ext_images/free.gif)
Salmonid species contain numerous short interspersed repetitive elements (SINEs), known collectively as the HpaI family, in their genomes. Amplification and successive integration of individual SINEs into the genomes have occurred during the evolution of salmonids. We reported previously a strategy for determining the phylogenetic relationships among the Pacific salmonids in which these SINEs were used as temporal landmarks of evolution. Here, we provide evidence for extensive genomic rearrangements that involved retropositions and deletions in a common ancestor of all the Pacific salmon and trout. Our results provide genetic support for the recent phylogenetic reassignment of steelhead and related species from the genus Salmo to the genus Oncorhynchus. Several other informative loci identified by insertions of HpaI SINEs have been isolated, and previously proposed branching orders of the Oncorhynchus species have been confirmed. The authenticity of our phylogenetic tree is supported both by the isolation of more than two informative loci per branching point and by the congruence of all our data, which suggest that the period between succesive speciations was sufficiently long for each SINE that had been amplified in the original species to become fixed in all individuals of that species. 相似文献
62.
Katsunori Kohda Yuji Tsuji Masahiro Takagi Tadayuki Imanaka 《Biotechnology letters》1996,18(9):1061-1066
Genes homologous to groES and groEL, which are recognized as molecular chaperone genes, from Bacillus stearothermophilus SIC1 were cloned and sequenced. By addition of GroES, GroEL and ATP in vitro, remaning activity of the alcohol dehydrogenase from Saccharomyces cerevisiae after heat treatment at 50°C for 6 min was improved from 55% to 90%. Furthermore, even though inclusion bodies were formed when a single chain Fv(sFv) was expressed in E. coli cells, during in vivo coexpression with molecular chaperone, a significant amount of the antibody protein could be recovered from the soluble fraction. 相似文献
63.
Tokuma Yanai Haruaki Taniguchi Hiroki Sakai Kazunori Yoshida Naoto Kimura Akira Katou Yuji Oishi Toshiaki Masegi 《Journal of medical primatology》1996,25(4):309-312
Abstract: A rare case of bilateral adrenal myelolipomas in a female cotton-top tamarin is reported. Large bilateral masses in the adrenal glands were composed of mature adipose cells containing varying amounts of hematopoietic cells of the myeloid, erythroid, and megakaryocyte series. The gross and histologic features of this case closely resemble human “giant” adrenal myelolipomas. 相似文献
64.
Binding of nonamer peptides to three HLA-B51 molecules which differ by a single amino acid substitution in the A-pocket 总被引:2,自引:2,他引:0
Akiko Kikuchi Takashi Sakaguchi Kiyoshi Miwa Yuji Takamiya Hans-Georg Rammensee Yutaro Kaneko Masafumi Takiguchi 《Immunogenetics》1996,43(5):268-276
The interaction between 9-mer peptides and HLA-B51 molecules was investigated by quantitative peptide binding assay using
RMA-S cell expressing human β2-microglobulin and HLA-B51 molecules. Of 147 chemically synthesized 9-mer peptides possessing
two anchor residues corresponding to the motif of HLA-B*5101 binding self-peptides, 27 paptides bound to HLA-B*5101 molecules. Pro and Ala at position 2 as well as Ile at position 9 were confirmed to be main anchor residues, while Gly
at position 2 as well as Val, Leu, and Met at position 9 were weak anchor residues for HLA-B*5101. The A-pocket is suspected to have a critical role in peptide binding to MHC class I molecules because this pocket corresponds
to the N-terminus of peptides and has a strong hydrogen bond formed by conserved Tyr residues. Further analysis of peptide
binding to HLA-B*5102 and B*5103 molecules showed that a single amino acid substitution of Tyor for His at residue 171(B*5102) and that of Gly for Trp at residue 167 (B*5103) has a minimum effect in HLA-B51-peptide binding. Since previous studies showed that some HLA-B51 alloreactive CTL clones
failed to kill the cells expressing HLA-B*5102 or HLA-B*5103, these results imply that the structural change of the A-pocket among HLA-B51 subtypes causes a critical conformational
change of the epitope for TCR recognition rather than influences the interaction between peptides and MHC class I molecules. 相似文献
65.
Selection of the best target site for ribozyme-mediated cleavage within a fusion gene for adenovirus E1A-associated 300 kDa protein (p300) and luciferase. 总被引:2,自引:1,他引:1
下载免费PDF全文
![点击此处可从《Nucleic acids research》网站下载免费的PDF全文](/ch/ext_images/free.gif)
H Kawasaki J Ohkawa N Tanishige K Yoshinari T Murata K K Yokoyama K Taira 《Nucleic acids research》1996,24(15):3010-3016
The cellular 300 kDa protein known as p300 is a target for the adenoviral E1A oncoprotein and it is thought to participate in prevention of the G0/G1 transition during the cell cycle, in activation of certain enhancers and in the stimulation of differentiation pathways. In order to determine the exact function of p300, as a first step we constructed a simple assay system for the selection of a potential target site of a hammerhead ribozyme in vivo. For the detection of ribozyme-mediated cleavage, we used a fusion gene (p300-luc) that consisted of the sequence encoding the N-terminal region of p300 and the gene for luciferase, as the reporter gene. We were also interested in the correlation of the GUX rule, for the triplet adjacent to the cleavage site, with ribozyme activity in vivo. Therefore, we selected five target sites that all included GUX The rank order of activities in vitro indeed followed the GUX rule; with respect to the kcat, a C residue as the third base (X) was the best, next came an A residue and a U residue was the worst (GUC > GUA > GUU). However, in vivo the tRNA(Val) promoter-driven ribozyme, targeted to a GUA located upstream of the initiation codon, had the highest inhibitory effect (96%) in HeLa S3 cells when the molar ratio of the DNA template for the target p300 RNA to that for the ribozyme was 1:4. Since the rank order of activities in vivo did not conform to the GUX rule, it is unlikely that the rate limiting step for cleavage of the p300-luc mRNA was the chemical step. This kind of ribozyme expression system should be extremely useful for elucidation of the function of p300 in vivo. 相似文献
66.
Gellan lyase was purified from the culture fluid of soil samples incubated in a medium containing gellan as a sole carbon source. The enzyme was a monomer with a molecular mass of 140 kDa and was most active at pH 7.5 and 45 degrees C. The enzyme was highly specific to gellan and lowered the viscosity of the polymer. 相似文献
67.
The immunomodulating effects of preoperative sizofiran (SPG) administration on regional lymph nodes were studied in patients
with stage III or IV head and neck cancer, by comparing the immunofunction of peripheral blood. The regional lymph nodes were
dissected surgically, and freshly obtained mononuclear cells were studied to investigate the interleukin-2 (IL-2) production,
the LAK and NK activities, and the quantitative analysis of the surface phenotype of the mononuclear cells. The results indicated
that SPG enhanced immunological activities in the regional lymph nodes, as shown by increased IL-2 production and cytotoxic
activities of the effector cells (NK, LAK), and increased helper T lymphocytes (CD4+) in the tumor-uninvolved lymph nodes. The immunofunction following SPG administration was attenuated, but was still augmented
in the regional lymph nodes with metastases. Therefore, SPG was found to be a biologic response modifier to enhance the immunofunctions
of the regional lymph node in patients with head and neck cancer. 相似文献
68.
The antitumor effect of the combined administration with recombinant human interleukin-2 (rIL-2) and sizofiran (SPG), a single
glucan of Shizophyllum commune Fries, was studied in vivo in C57BL/6 mice intraperitoneally inoculated with EL-4 lymphoma.
The effect was evaluated by a) comparing the survival time of the mice, b) analysis of the intraperitoneal cell population
in Giemsa-stained specimens, c) surface marker analysis of peritoneal exudative cells with flow cytometry, d) cytotoxic assay
of cells against EL-4 and Yac-1 lymphoma, and e) elimination of some cell populations by monoclonal antibodies, to identify
the antitumor-effector cells showing cytotoxic activity. The survival of mice given both rIL-2 and SPG was significantly longer
than the control mice or those given SPG alone or rIL-2 alone. It was demonstrated that the administration of SPG and/or rIL-2
to the EL-4 lymphoma-bearing mice activated immune-response cells in the peritoneal cavity such as T lymphocytes, NK cells,
or macrophages, which might be effective in reducing lymphoma cells. The combination of rIL-2 and SPG administration appears
to activate the antitumor- immune response at the tumor site more effectively than when either agent was administered alone. 相似文献
69.
Hisakazu Mihara Kin-Ya Tomizaki Norikazu Nishino Tsutomu Fujimoto Haruhiko Tamaoki Yuji Kobayashi 《Biopolymers》1994,34(7):963-967
A model 16-peptide of endothelin-1 (MET-1), which has the minimized sequence homology to the corresponding pan of endothelin-1 (ET-1), was designed to confirm the cystine-stabilized α-helix motif. The model structure consists of an extended structure, a β-turn part, and an α-helix structure that is stabilized by two disulfide bonds. The α-helix segment was designed to emphasize the amphiphilic nature. In order to combine the extended structure and the α-helix segment, a D -Ala-Pro sequence was selected to fix the β-turn. The model endothelin 16-peptide amide was synthesized by solid-phase synthesis on a 4-methylbenzhydrylamine resin. Its conformation was examined by CD and two-dimensional (2D) 1H-nmr measurements. MET-1 showed similar CD patterns to ET-1 in both buffer and 50% aqueous trifluoroethanol solution. The 2D nmr experiments in 50% aqueous ethylene glycol revealed that MET-1 closely resembles the conformation of ET-1 with an extended structure, an α-helix, and a β-turn unit in the same position of the sequence. Furthermore, model peptides without disulfide bond(s) could not assume a stable structure in aqueous solution, while they did have similar α-helical content in 50% trifluoroethanol with MET-1. When the two disulfide bridges were simultaneously formed, the peptide with the correct disulfide bonds (MET-1) was obtained in threefold excess to the isomer (apamin type. MET-2). These findings obtained by the modeling of ET-1 showed an important role for the stabilization of peptide conformation with disulfide bonds. © 1994 John Wiley & Sons, Inc. 相似文献
70.
Mitotic chromosomes of the plant pathogenic filamentous fungi Botrytis cinerea and Alternaria alternata were observed. Chromosomes prepared by the germ tube burst method were stained with the fluorescent dye 4,6-diamidino-2-phenylindole (DAPI) to yield figures with good resolution. Using this method, component chromosomes were clearly distinguished and the chromosome number could be determined. Fluorescence in situ hybridization (FISH) was also successfully applied to the specimens, revealing one ribosomal RNA gene cluster, or nucleolus organizer region (NOR) in the genome of each fungus. A long attenuated chromatid thread expanding from a condensed metaphase chromosome, which had been called a thread-like structure in B. cinerea, was proved to be an NOR. This is the first report of the successful application of FISH to the chromosomes of filamentous fungi. 相似文献